11P WU-NK-101: an Enhanced NK Cell Therapy Optimized for Function in the Tumor Microenvironment (TME)

The efficacy of adoptive cell therapies against solid tumors has been limited by the immune-suppressive TME, which aids immune escape. WU-NK-101, manufactured using the MONETATM platform, is an NK product derived from healthy donor NK cells that addresses challenges of adoptive cell therapy in the setting of an adverse TME. Cell phenotypes were evaluated by mass cytometry. Cytotoxicity was assessed in vitro in 2 culture conditions: IMDM-20 and TME-aligned media. Multi-omic studies were performed by mass spectrometry. Cell trafficking/penetration to the TME was measured in NSG mice by tracking labeled WU-NK-101 cells ± mAb. WU-NK-101 has a unique phenotype vs. conventional NK cells (cNK), with higher expression of activating receptors, which enable rapid activation and improved activity. The latter was confirmed by cytotoxicity assays, with lower Effector:Target (E:T) ratio required to kill 50% of target cells (EC50) for WU-NK-101 vs. cNK, mEC50 1.7 vs. 5.2 (p= 0.032). Importantly, TME-aligned media had a strong impact on cNK killing but negligible impact on WU-NK-101 cytotoxicity, mEC50 16.3 vs. 2.0 (p= 0.018). Potentially explaining this improved killing, WU-NK-101 has metabolic profile consistent with aerobic glycolysis, which may counteract the inhibition of the TME. In IMDM-20, WU-NK-101 uses glucose as its main nutritional source; while in hypoglycemic TME-aligned media, amino acid metabolic pathways were upregulated: this metabolic adaptability augurs improved in vivo efficacy in solid tumor microenvironments. To confirm anti-tumor activity in mouse models, WU-NK-101 was able to infiltrate tumors with robust persistence and anti-tumor activity, which was enhanced with mAb pre-dosing (p = 0.016). WU-NK-101 has potent cytotoxicity against tumor cells and enhanced/adaptive metabolic fitness contributing to resilience toward the immunosuppressive TME, relative to cNK cells. mAb addition further enhances anti-tumor activity by improved NK trafficking and tumor penetration, taking advantage of these enhanced metabolic features of WU-NK-101. These data suggest that WU-NK-101 may overcome the limitations of adoptive cellular therapy and supports clinical development in the setting of solid tumor.